Cell Culture

Boyden chambers assay is a useful tool to study cell migration and cell invasion such as chemotaxis, haptotaxis and transmigration. It consists of a cylindrical cell culture insert nested inside the well of a cell culture plate.

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Clonogenic assay or colony formation assay is an in vitro cell survival assay based on the ability of a single cell to grow into a colony. It is the method of choice to determine cell reproductive death after treatment.

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sproutingThe spheroid sprouting assay consists of a self-aggregation of lymphatic endothelial cells (LECs) embedded in a 3D matrix. This system allows to determine the area occupied by the migrated cells and their spatial distribution.

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tubulogenesisTubulogenesis assay consists of lymphatic endothelial cells (LECs) cultured between two layers of type I collagen. This systems allows to determine the area density of capillary-like structures formed by LECs.

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lymphatic ring assayThis system consists of ex vivo culture of fragments of thoracic ducts dissected from mice. This assay allows to study the sprouting of lymphatic capillaries from an authentic lymphatic vessel.

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This assay allows the assessment of the migratory capability of lymphatic endothelial cells (LECs).

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Immunosuppression can be evaluated by cell cycle analysis of PBMC stimulated (or not) with anti-αCD3/CD28 microbeads and cultivated during 4 days with or without test cells (MSC…). Cell cycle analysis is performed using the CycleTEST Plus DNA Reagent Kit and acquisition by flow cytometry. Immunosuppressive properties of cells are quantified by their inhibition of stimulated PBMC-induced proliferation.

The Flexercell ® FX4000™ is a computer-regulated bioreactor that uses vacuum pressure to apply cyclic or static strain to cells, cultured on flexible-bottomed culture plates. The force exerted and its frequency can be modulated. This model allows the study of traction on the metabolism of cells. It helps to study the effects of mechanical forces of various intensities and frequencies on the phenotype of adherent cultured cells. Moreover, different types of forces (tension, shear stress, compression) can be applied in 2D or 3D models.

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The rotary cell culture system is a device designed to grow 3D cells clusters (tissues, cancer tumors and virus cultures) in microgravity (outside the body). The advantage here is to obtain larger and 3D cell cultures with structural and chemical characteristics similar to normal tissue, in comparison with lab-grown cell cultures.

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We are equipped with GammaCell® 40 EXACTOR. It is ideal for the irradiation of mitotically inactive cell culture media, mixed lymphocyte cultures and cellular blood components. Redundant systems monitor the essential irradiation parameters, time and position of sources. Dose Rate and Radiation Specifications:

  • Central dose rate of approximately 1.0 Gy/minute (100 rad/minute)
  • Each of the two special form Caesium-137 sources has a nominal activity of 55.5 TBq (1500 Ci). Together they produce a central dose rate of 1.0 Gy/minute (100 rad/minute) ±15% in the sample container.

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An attractive feature of human induced pluripotent stem cells (also known as iPS cells or iPSCs) is the ability to derive them from adult patients to study the cellular basis of human disease. Since iPS cells are self-renewing and pluripotent, they represent a theoretically unlimited source of patient-derived cells which can be turned into any type of cell in the body.

The researchers at GIGA developed the expertise in deriving iPS from skin fibroblasts by introducing a specific set of pluripotency-associated genes.

Upon introduction of reprogramming factors, cells begin to form colonies that resemble pluripotent stem cells which can be isolated based on their morphology, conditions that select for their growth, or through expression of surface markers or reporter genes.

iPSCs are capable of differentiation in a fashion similar to Embryonic Stem Cells into fully differentiated tissues.

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The Hematology unit of the CHU and several teams at GIGA are specialized in the study of hematopoietic and other stem cells (e.g. mesenchymal stem cells, neural crest stem cells).

This includes investigations of stem cell biology, purification and expansion, as well as the therapeutic use of these cells in the context of hematopoietic cell transplantation and solid organ transplantation, immunotherapy of cancer, auto-immune diseases and regenerative medicine.

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The dermal equivalent is an in vitro model of the dermal layer of skin. It is constructed by seeding dermal fibroblasts into a collagen gel. Other cell types may be incorporated into the dermal equivalent to increase the complexity of the model.

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Human keratinocytes and melanocytes cultured in vitro are grown on a biological matrix (dead de-epidermized human dermis) and the system is kept at an air-liquid interface, in a suitable culturing medium, until a stratified human epidermis is formed, maintaining the histological characteristics of the epidermis in vivo. De-epidermized dermis model is a closer approximation to the in vivo situation of formation and maintenance of the mature epidermis.

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Our researchers at the ULg have developed an innovative chitosan-based scaffold for wound repair and tissue engineering. This new scaffold-type has unique structural properties that stimulate the healing process and improve of the overall quality of the repaired tissue through a more adequate revascularization, re-epithelialization and remodeling of the granulation tissue. Easy-to-handle, and large potential for re-modelling.

For more info, click here

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Health Economics

Economic evaluation has become increasingly important to inform decision makers about how to allocate scare resources, especially with the increasing health care expenditures, the rapid introduction of new medical technologies and the extending role of economic evaluations in health care decision making. Medical innovations are nowadays introduced and diffused in a controlled manner and have to respond to economic criteria. We aim to evaluate the economic value of innovative medical technologies in order to help public and private decision makers to efficiently allocate scare health resources. Our multidisciplinary team provides services, analyses and expertise for several academic, public and private decision makers to efficiently allocate scare health resources, by estimating the relative efficiency of therapeutic and diagnostic interventions (e.g. cost-effectiveness analyses) and by using more innovative methods (e.g. discrete choice experiments, risk-sharing models).

Systematic Reviews & Meta-analyses

Systematic reviews and meta-analyses are key elements of Evidence Based Medicine and Evidence Based Health Care. We have a strong experience in the design and the methodology of systematic reviews and meta-analyses. We use explicit and transparent methods following a standard set of stages and our results are accountable, replicable and updatable. We use the PRISMA statement, which is an Evidence Based minimum set of items for reporting in systematic reviews and meta-analyses. We also use the AMSTAR check list to check the methodological quality of our systematic reviews.

Biostatistics

Excellence in biostatistics is a core strength at CHU/ULg. Our talented biostatisticians offer a depth of knowledge and experience in all aspects of clinical research from study design, document preparation for ethic committee review, data analysis, results interpretation to final report.

Our biostatistical expertise has been sharpened over the year in collaboration with biomedical researchers, clinical physicians, public health institutions and pharmaceutical companies. It covers the following domains:

 

  • Study design and implementation (with support of epidemiologists, clinicians and other experts)
  • Project protocol finalization
  • Sample selection, design of data collection instruments, data quality control and data management

More specifically, they help to prepare this document thanks to the following services:

  • Cross-over, sequential, group-sequential, adaptive designs, seamless phase II/III
  • Large, simple trials
  • Patient selection (wide versus restricted population)
  • Control of heterogeneity (stratification)
  • Treatment allocation methods (minimization, permuted blocks, etc.)
  • Scales to measure and standardize treatment benefits or harm
  • Sample size and power calculation
  • Duration and intensity of follow-up
  • Factorial and multivariate designs
  • Methods of analysis (including interim analyses)
  • Pharmacokinetic and pharmacodynamic (PK/PD) modeling
  • Bioequivalence and bioavailability studies
  • Evaluation of the maximum tolerated dose

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  • Basic statistical analysis
  • Statistics on rates and proportions
  • Parametric and non-parametric tests
  • Linear and non-linear regression models
  • Longitudinal/correlated data analysis
  • Survival analysis
  • Multivariate methods
  • Censored and missing data
  • Bayesian models
  • Big data

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Clinical data are presented with a statistical analysis and complete appendices in an integrated report. At your convenience the final integrated report is delivered in PDF format or as hard copy. Furthermore, from your raw data, we are able to produce a publishable summary of the project findings

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The team is familiar with the following statistical packages:

  • SAS: software system for data analysis and report writing. This program allows to store data values and retrieve them, modify data, compute simple and complex statistical analyses and create reports
  • R: software system object-oriented for statistical computing and graphics
  • S-PLUS: software system object-oriented programming capabilities and advanced analytical algorithms
  • JMP: software system used in applications such as Six Sigma, quality control and engineering, design of experiments and scientific research. The software is focused on exploratory analytics, whereby users investigate and explore data, rather than testing a hypothesis
  • WinBUGS: software system used for Bayesian analysis using Markov chain Monte Carlo (MCMC) method. It is based on the BUGS (Bayesian inference Using Gibbs Sampling).
  • Epi-Info: public domain statistical software for epidemiology, allowing electronic survey creation, data entry and analysis. Within the analysis module, analytic routines as well as analysis of complex survey data are available

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Researchers and investigators are also encouraged to consult computer science specialists in the planning stage of their project. Our computing team has a long-standing expertise in offering support for tailored public health and medical computer applications. Specifically, it covers the following domains:

  • Design and specifications of informatics projects
  • On-line surveys and questionnaires
  • Development of web-based applications
  • Innovative creation of websites (ASP.NET)
  • Database management (SQL Server / MySQL)
  • Database storage
  • Large-scale screening surveys

Regarding safety, the University Hospital offers an optimal security system for the storage of electronic data. The system is subject to regular revisions, both physically and technologically. Moreover, the data security system is approved by the Sectoral Committee of the National Register. The University Hospital has a data security policy as well as a “Code of Ethics and Patient Data Secure”. This document is made available to the Committee.

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Cell therapy

The CHU Laboratory of Cell and Gene Therapy (LTCG) has developed a strong expertise in basic research on in vitro and murine models, in the production of cellular product batches (ATMP) and in running related phase I/II clinical trials.

The LTCG consists of:

  • 3 FAMHP-approved tissue banks (hematopoietic cells, cord blood, non-hematopoietic cells) and a GMP facility offering services like cell collection (from patients or normal volunteers), processing, storage and distribution
  • R&D unit to transfer innovative cell technologies to the clinic
  • ATMP production lab producing and distributing cell products used in patients in the context of approved clinical trials

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  • Delivering of human HSC, PBMC, cultured BM or cord MSC, etc
  • Minimal processing of cellular products (red cell removal, cell selection and depletion, freezing & thawing)
  • Production of cellular ATMP
  • Immunotoxicity tests (humanized mouse models in NOD/SCID/IL-2Rγ(null) mice)
  • Clean room rental
  • Consulting
  • Phase I-II clinical trials
  • The CROPHA accreditation
  • The FACT-Netcord
  • The JACIE (joint accreditation committee ISCT-EBMT)

Biobank

Biobank_NV site

Our biobank offers a collection of human biological samples (frozen or embedded in paraffin, pathological or non-pathological) with relevant clinical and technical data respecting all ethical, legal and quality requirements.

In addition, our FAMHP-approved tissue bank has a large number of samples of hematopoietic cells, cord blood and non-hematopoietic cells.

Should you request samples, contact us (info@b2h.be), we will help you to submit your project to the Ethical Committee to get approval and grant you access to the samples.

 

Viral vectors

Logo_Logo-autres_GIGAThe GIGA viral vector platform has a large expertise in the production of retro/lentiviral/AAV vectors and the generation of stable cell lines.

Our retro/lentiviral vectors (2nd or 3rd generation systems) contain all necessary biosafety features.

Services offered:

  • Viral vector design fitting your specific requirements
  • Ready-to-use viral vector systems (with different pseudotypes/serotypes and promoters) carrying fluorescent proteins
  • Support and advice to define best suited viral vectors for optimal cell transduction

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  • Maxiprep using PureYield™ Plasmid Maxiprep System (from Promega)
  • Endotoxin, RNA, protein removal from purified plasmid DNA
  • Plasmid transformation into bacteria

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Viral vector production, concentration and titration for:

  • Custom Adeno-associated viral vectors (rAAV): different serotypes are available (AAV2/1, AAV2/2, AAV2/5, AAV2/8, AAV2/8, AAV2/DJ) and other serotypes on demand
  • Custom and pre-made retro Lentiviral vectors (rLV) derived from HV-1: depending on the target cells, different pseudotypes (VSV-G, Ampho, Eco) can be used

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Cell transduction, cell selection and tests of the supernatant regarding the absence of replication competent virus and mycoplasma:

  • Over-express your gene of interest: the platform has some premade viral vector plasmids and viral backbone plasmids
  • Knockdown expression of your gene of interest: shRNA plasmids viral vectors (Sigma shRNA Mission)

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CRISPR-Cas9 is an RNA-guided genome editing system, which can be used for gene disruption, gene knock-out, gene mutations and controlled gene insertion.

We are able to produce lentiviral vectors expressing Cas9 and RNA guide for targeting your gene of interest.

We have established a partnership with Sigma Aldrich and are members of Sigma CRISPR Core.

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